Operating the JEOL JEM1400-Plus Transmission Electron Microscope

Electron Microscopy Core Laboratory, University of Utah

Follow the procedures below when you use the JEM1400-Plus TEM.

Starting Session
1. If not logged in, log in to the "user" accounts on the JEOL and Gatan computers. (Use keyboard and mouse
lying on top of Gatan computer to log into that computer.) After this, the computers should be controllable by
JEOL mouse and keyboard. If not, check that "Synergy" is running on both machines.
2. Check that the gun and column vacuum is reasonable (Penning Gauge < 30 and "Vac Ready", other chambers
except "specimen chamber" should say "Evac Ready"). Use "valve/vacuum monitor" panel.
3. If the "high tension" is off, turn it on. It should be set to 120 kV (lower if a different value is desired).
-Use "beam controller"
-If high tension shuts off as you bring up the voltage, raise voltage to a point lower than where the high tension
failed. Let voltage stay there for a few minutes. Then, bring up high tension slowly. For example, turn
voltage to 100 kV with "beam controller", then use "HT Scheduling" to go from 100120 kV in 0.1 kV steps of
one second.
4. Add liquid nitrogen to the cold trap.
-Remove "ACD" (anti-contamination device) attachment if present.
-Test if connections are okay by pushing (up) switch at top of dewar (below amber light). If amber light
changes color, then don't add nitrogen and notify EM Lab staff. This indicates a conductivity problem with the
microscope. If this occurs when you have added nitrogen, also notify staff.
-Use a funnel to add nitrogen.
-Add more nitrogen during your session as needed (e.g. hourly or during specimen exchanges).
5. Aperture settings
-condenser aperture should be IN (adjust with lever; left is in, right is out).
-objective aperture should be OUT (red dot is out, white dots are in and show relative size of aperture). The
objective aperture may be IN after you know you have a beam and if you want the extra contrast.
-selected area (SA) aperture should be OUT (adjust with lever; left is in, right is out). As with the objective
aperture, if you have a beam, the SA aperture may also be IN during use if you wish to restrict the view to
specific regions of your specimen.

Viewing Specimen
1. Insert specimen (see instructions below)
2. After the "Penning Gauge" vacuum reaches 30 or less and the specimen chamber says "evac ready", turn on
filament. (Filament is on when "On" and "Beam Ready" buttons turn bright green.)
3. Check for beam. If no beam, then see instructions "finding the beam" (on separate sheet).
4. Set eucentric position, i.e. move stage to the z=0 position. Either of the following can work:
a. Rotate the stage to get minimum shift
-Set magnification to a value in the range 20005000
-Find a feature on the specimen and center that feature on the fluorescent screen
-Rotate the stage (~20 is usually sufficient) by pressing the "+TX" and "TX" buttons in the "Stage
Controller" panel.
-Observe the feature as the rotation occurs. Does the feature move?
-If the feature moves, adjust z-position by pressing "+Z" or "Z" button in the "Stage Controller" panel.
-Repeat the last three steps until the movement of the feature during the rotation is minimized. The
observed specimen is now at or near the eucentric position.
b. Adjust z to focus position
-Press "Standard Focus" button on right console
-Move z to focus position by using the "+Z" and "Z" buttons in the "Stage Controller" panel.
-Alternatively, adjust the z-axis position by pressing "Z focus" button (right panel) and then turning the
"Focus" knob. (Note: if "Z focus" is not pressed, the z-axis position will not be changed and focusing
occurs by changing lens currents.)
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c. Adjust to new eucentric height when you move to a new part of the grid (e.g. when you move to a different
grid square or a significant change in x or y).
Focusing: use the following ideas to focus.
a. Ways to adjust focus
-Adjust focus knob (right console).
-Focus using adjustment of z-axis (see above)
-Press "Standard Focus" button on right console. If your specimen is in the eucentric position, you should
now be reasonably close to the focus position. However, you likely will need to use another method to
find the exact focus point.
-Wobbler button ("IMG WOB") is also on right console. Press this button and then adjust focus. This
method can be especially helpful for focusing at lower magnifications.
b. Recognizing focus
-If you use the wobbler, adjust focus until the image wobble is minimized. You will be at or near the focus
point.
-Find minimum contrast on fluorescent screen or the Gatan camera. Edges are usually the easiest place to
find the minimum contrast point.
-Using the FFT option in Gatan Digital Micrograph, find focus as the setting between under-and over-focus
Thon rings. At the focus point, the Thon rings disappear. A square FFT will work best. (Otherwise, the
rectangular nature of the Orius camera will produce a rectangular FFT, and the FFT may give the false
impression of astigmatism.) To obtain a square FFT in DM, select region in micrograph using
"rectangular region of interest" option (dashed square in symbol menu). Hold down "Alt" key to select
square. Then select FFT (live or still). If Thon rings are difficult to see, move to a different part of the
specimen or adjust the histogram.
-Often, a combination of these methods is necessary to find focus.
Align microscope, see alignment instructions
Record images (see instructions below)
Get help if something is amiss. Notify EM Core Lab staff if problems arise (e.g. column dumps).
Hints
-"Beam Blank" (is one of the left buttons on the table console): use to blank beam
-"Nanospace Map" (under "Control", "Stage"): use it to move to specific areas on the stage (select point, then
click "Shoot")
-"Image Forming System Controller" can be used to set multiple magnification settings that you can quickly
move to by clicking the "MAG2" or "Low MAG" buttons. (Use "Low MAG" for lower magnifications, e.g.
1000x or less.)
-Push console buttons to switch between fine and coarse steps. Coarse or fine is indicated by the light adjacent
to the button. The coarse setting has a brighter light.
-Console buttons will to "sleep" if left unused for a period of time. Push any button to "awake".

Insert Specimen
1. Load specimen onto specimen holder
a. single specimen holder
-use tweezers to open clip
-place specimen grid in holder
-use tweezers to clamp clip on specimen
-tap holder to ensure specimen is firmly in place
b. 5-specimen holder
-for each specimen position, do the following two things
-place specimen grid in holder
-place holder ring above specimen and screw in place with special tool
2. Insert specimen holder into microscope (see p. 5-9 of JEM-1400 manual)
*If inserting the 5-specimen holder, BE SURE SPECIMEN 5 IS IN THE VIEWING POSITION. HOLDER
COULD BE DAMAGED IF NOT.
-place holder into TEM, align guide pin with guide groove, push in gently without turning until the holder stops
-turn switch from "Air" to "Pump", you must pull switch out and push it up
-wait for green light to come on
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-After green light comes on, rotate clockwise a few degrees until it stops and is drawn into the microscope,
specimen holder will be drawn into the microscope a short distance until it stops (don't let the holder slam into
the next position)
-rotate clockwise several degrees until holder stops, specimen holder will be drawn completely into the
microscope (don't let the holder slam into position)
Select appropriate holder in "Specimen" panel.

Remove Specimen
1. Turn filament off.
2. Neutral stage (stage moved to XY center and =0), use "Stage Controller" panel. ("Exchange Holder" button
will neutralize stage and turn off filament.)
3. Retract Gatan camera
4. Put fluorescent screen down
5. If removing the 5-specimen holder, BE SURE SPECIMEN 5 IS IN THE VIEWING POSITION. HOLDER
COULD BE DAMAGED IF NOT.
6. Remove holder
-pull sample holder straight out until it stops
-turn holder anti-clockwise several degrees until it stops, if you feel resistance, STOP and get help, something is
amiss
-pull holder out a short distance until it stops
-turn holder a few degrees anti-clockwise until it stops
-turn switch from "Pump" to "Air", you must pull switch out and push it down
-wait for amber and green and lights to turn off and hissing sound
-pull holder out of microscope
7. Remove grid from holder
a. single specimen holder
-use tweezers to open clip
-remove grid
b. 5-specimen holder
-unscrew holder ring
-remove grid

Alignments
1. If desired, reset to default alignments under "Alignment" panel. Click "Neutralize Selected Alignment".
2. For typical user, do all alignments at magnification you will use to record your images (highest if you will use a
range of magnifications). However, align at least in the range 2000 to 5000. For JEOL (supervisor) account,
use 10,000 or higher.
3. Center condenser aperture
-condense beam to a spot with "BRIGHTNESS" knob
-move beam to center of fluorescent screen with beam shift ("X", "Y") knobs
-expand beam with "BRIGHTNESS" knob
-when centered, beam will expand symmetrically around the fluorescent screen
-adjust knobs on condenser aperture until expansion is symmetrical
4. Open "Alignment" panel
5. At any time, adjust condenser stigmation by pressing "COND STIG" button
6. Adjust gun tilt
-select "GUN Tilt"
-open "Film Camera" and observe exposure time
-minimize exposure time by adjusting "X" and "Y" knobs
7. Adjust gun shift
-select "GUN Shift"
-select spot size 1
-condense beam to smallest size with "BRIGHTNESS" knob
-move beam to center of fluorescent screen with "X" and "Y" knobs
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-select "Beam Shift"

-select spot size 4
-condense beam to smallest size with "BRIGHTNESS" knob
-move beam to center of fluorescent screen with "X" and "Y" knobs
-repeat last eight steps until beam remains at center of screen
Align beam tilt
-focus image, be sure you have done at least a rough focus in Z (to set specimen at eucentric height)
-select "Beam Tilt"
-open "Wobbler" controller
-select "Object Lens" on wobbler panel
-minimize shift of image with "X" and "Y" knobs (Hint: do one knob at a time and minimize movement. You
want to have rotation around the center or a central feature makes a small circle. The edges rotate.)
Adjust objective lens stigmation
-use Gatan camera
-select "OBJ STIG" in "Alignment Panel", click again when done
-record image (live or still)
-make Thon rings round (see focusing section under "Viewing Specimen" above)

Record Images with Gatan Camera

1. Turn on controller if not already on. "Cooler" light will be orange until correct temperature (10C) is reached
(data light will flash as data is collected).
2. Turn on Digital Micrograph (DM) using menu on Gatan computer screen, you should hear two beeps
3. Insert camera with option in DM (microscope vacuum sucks camera in, compressed air pushes it out)
4. Press "Start View" to run live camera (images usually binned by 24)
5. Press "Start Acquire" to record full size image (images usually unbinned)
6. Save your images, if desired
-Images may be saved to computer hard drive (drive D only), but may be deleted after one month. If saved to
drive D, save in the following manner: Initial folder is the PI name (LastName_FirstName). Thereafter, save
in folder with the name of user (in format LastName_FirstName). The date (YEAR-MONTH-DATE in
numerical format XXXX-XX-XX) should be included in a subfolder. Project or sample name may also be
included, but include the date in at least one subfolder name.
-Save images to flash drive plugged into Gatan computer
-DO NOT install any software (including drivers) on the Gatan computer. If your drive will not work without
installing drivers on the Gatan computer, then find another drive that will.
-DO NOT plug anything into JEOL computer
7. Hints
-0.0625 sec exposure may be an appropriate setting for "search" mode, a longer time may be useful especially
when using live FFT.
-"auto survey" option will allow automatic light normalization
-Batch convert: use "save display as" option.

Ending Session
1. Set magnification to 2000
2. Spread beam to cover approximately half of the screen, center beam if uncentered.
3. Turn off filament
4. Remove specimen (follow instructions above)
5. IF YOU ARE THE LAST USER OF THE DAY, DO THE FOLLOWING:
-attach ACD (anti-contamination device), including plug
-open all apertures (condenser, objective, and SA) by turning lever to right
-turn off high tension
-turn on ACD heat (click "Control", "Maintenance", "ACD" to get ACD controller)
by David Belnap, August 22, 2014
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